mouse cxcl12  (Proteintech)

Journal: Non-coding RNA Research

Article Title: Aerobic exercise alleviates allergic airway inflammation by suppressing circMETTL9 -mediated formation of macrophage extracellular traps ⋆

doi: 10.1016/j.ncrna.2025.08.008

Figure Lengend Snippet: Communication Between Airway Epithelial Cells and Macrophages Mediated by CXCL12-CXCR4 Regulates METs. (A) The co-expression network of IGFBP3 and macrophage chemokines was predicted by the STRING database. (B) The binding of CXCL12 to the CXCR4 receptor was predicted in the CellphoneDB database. (C) Representative SYTOX Green staining in macrophages treated with or without CXCL12 (n = 3). (D) The protein expression of MPO and CitH3 in macrophages was detected by Western blot analysis (n = 3). (E – G) ELISA was used to detect the expression of CXCL12 in the cell supernatant of the co-culture system (n = 3). RT-qPCR was used to detect the expression of CXCL12 in BEAS-2B cells and CXCR4 in macrophages (n = 3). All data are expressed as means ± SD. ∗ P < 0.05. GAPDH was used as a loading control for all Western blot assays. All data are expressed as means ± SD. ∗ P < 0.05.

Article Snippet: To assess the expression levels of CXCL12, ELISA kits specifically for mouse CXCL12 (catalog #KE10049, Proteintech, Chicago, USA), and for human CXCL12 ELISA Kit (catalog #RK00266, ABclonal, China) were utilized according to the manufacturer's protocols.

Techniques: Expressing, Binding Assay, Staining, Western Blot, Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Quantitative RT-PCR, Control

Journal: Non-coding RNA Research

Article Title: Aerobic exercise alleviates allergic airway inflammation by suppressing circMETTL9 -mediated formation of macrophage extracellular traps ⋆

doi: 10.1016/j.ncrna.2025.08.008

Figure Lengend Snippet: Overexpression of CircMETTL9 Counteracts the Reduction Effect of Aerobic Exercise on METs. (A) Schematic timeline of the experimental protocol. Day 0: AAV-LUNG-OE- circMETTL9 by the nasal drip. Days 14, 28, 42, and 56 represent intraperitoneal (i.p.) injections of OVA. Days 35–68 represent exposure to ovalbumin aerosol. Aerobic exercise adaptation occurred from days 35–37, and days 39 and 67 represent the initial and final physical tests. Aerobic exercise was initiated on day 42 and ended on day 66. Euthanasia was performed on day 70. (B) The expression of circMETTL9 was detected by RNA FISH staining (n = 6). (C) The expression of circMETTL9 was performed by RT-qPCR (n = 6). (D) ELISA was used to detect the expression of CXCL12 in the BALF (n = 6). (E) The mRNA expression of CXCL12 and CXCR4 was detected by RT-qPCR (n = 6). (F) Western blot analysis was used to detect the protein expression of CitH3 and MPO in the lung tissue (n = 6). (G) Representative immunofluorescence images of CitH3, MPO, and CD68 staining of lung tissues. (H) The result of the Western blot showed the effect of circMETTL9 overexpression on IGFBP3 and EIF4A3 expression (n = 6). A, OVA-induced asthmatic mice and infected with blank AAV; E, mice were subjected to aerobic exercise and infected with blank AAV; OE-A, OVA-induced asthmatic mice and infected with AAV overexpressing circMETTL9 ; OE-AE, OVA-induced asthmatic mice performed aerobic exercise and infected with AAV overexpressing circMETTL9 . All data were shown as the means ± SDs and were assessed by a paired two-tailed t -test. ∗ P < 0.05.

Article Snippet: To assess the expression levels of CXCL12, ELISA kits specifically for mouse CXCL12 (catalog #KE10049, Proteintech, Chicago, USA), and for human CXCL12 ELISA Kit (catalog #RK00266, ABclonal, China) were utilized according to the manufacturer's protocols.

Techniques: Over Expression, Aerosol, Expressing, Staining, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Western Blot, Immunofluorescence, Infection, Two Tailed Test

Journal: Journal of Biomedical Research

Article Title: DEC1 promotes breast cancer bone metastasis through transcriptional activation of CXCR4

doi: 10.7555/JBR.39.20250031

Figure Lengend Snippet: DEC1 deficiency prevented breast bone metastasis induced by intracardiac injections of 4T1 cells via decreasing CXCR4/CXCL12 in mice. Forty 4-month-old mice, comprising twenty Dec1 +/+ and twenty Dec1 −/− , were divided into four groups: Dec1 +/+ -PBS, Dec1 +/+ -4T1, Dec1 −/− -PBS, and Dec1 −/− -4T1. Mice in the Dec1 +/+ -4T1 and Dec1 −/− -4T1 groups received an intracardiac injection of 4T1 mouse breast cancer cells to induce breast bone metastasis, while mice in the Dec1 +/+ -PBS and Dec1 −/− -PBS groups received an equal volume of PBS via the same method for two months. n = 10 for each group. A: Osteolytic bone injury in the four groups of mice by X-ray. B: The serum CXCL12 amount in the four groups of mice ( n = 3–5). C–F: The CA153, CK8, and Ki67 expression in the four groups of mice by immunohistochemical staining ( n = 3). G and H: The TRAP-positive cells in the four groups of mice ( n = 3). I–N: The protein levels of CXCR4, TRAP, N-cadherin, E-cadherin, and vimentin in the four groups of mice by Western blotting ( n = 3). O: The CXCR4 expression in the four groups of mice. Data are presented as mean ± standard deviation. * P < 0.05, ** P < 0.01, *** P < 0.001, and ns P > 0.05, comparisons are shown in the figure. Data were analyzed using two-way ANOVA, and differences between groups were analyzed using Student's t -test.

Article Snippet: CXCL12 enzyme-linked immunosorbent assay (ELISA) Kit (Cat. #KE10046) was from Proteintech (Wuhan, China).

Techniques: Injection, Expressing, Immunohistochemical staining, Staining, Western Blot, Standard Deviation

Journal: Journal of Biomedical Research

Article Title: DEC1 promotes breast cancer bone metastasis through transcriptional activation of CXCR4

doi: 10.7555/JBR.39.20250031

Figure Lengend Snippet: DEC1 promoted CXCL12 production from mesenchymal stromal cells in mice and MC3T3-E1 cells. A: The CXCL12 expression in the femur of wildtype (WT) and Dec1 -knockout (KO) mice by immunohistochemical staining ( n = 3 in each group). B: The relative Cxcl12 mRNA levels in bone tissues from the femur of WT and Dec1 -KO mice ( n = 5 in each group). C: The relative Cxcl12 mRNA levels in mesenchymal cells from the femur of WT and Dec1 -KO mice ( n = 3 in each group). D–F: MC3T3-E1 cells were seeded into 6-well plates and cultured overnight. Cells were divided into three groups: Vector (infected with LV-shCon and transfected with Flag-CMV2), DEC1 -OE (transfected with Flag- DEC1 ), and DEC1 -KD (infected with LV-sh DEC1 ). After 24 h of transfection or infection, the culture medium from each group was collected, centrifuged, and filtered to obtain conditioned medium (CM). Effect of CM from MC3T3-E1 cells with DEC1 -OE or KD on breast cancer cell migration by Transwell migration assays (D). The relative Cxcl12 mRNA levels in MC3T3-E1 cells (E). The CXCL12 amount in CMs from DEC1 -OE and KO in MC3T3-E1 cells by ELISA assays (F). G: Mechanism of DEC1 promoting breast cancer bone metastasis via transcriptional activation of CXCR4. Data are presented as mean ± standard deviation (all experiments were repeated at least three times). * P < 0.05, ** P < 0.01, *** P < 0.001, comparisons are shown in the figure. Data were analyzed using one-way ANOVA, and differences between groups were analyzed using Student's t -test. Abbreviation: EMT, epithelial-to-mesenchymal transition; KD, knockdown; OE, overexpression.

Article Snippet: CXCL12 enzyme-linked immunosorbent assay (ELISA) Kit (Cat. #KE10046) was from Proteintech (Wuhan, China).

Techniques: Expressing, Knock-Out, Immunohistochemical staining, Staining, Cell Culture, Plasmid Preparation, Infection, Transfection, Migration, Enzyme-linked Immunosorbent Assay, Activation Assay, Standard Deviation, Knockdown, Over Expression